Carbohydrates

Qualitative Analysis
of Carbohydrates
Dr Tahira Leghari
Biochemistry Department
Carbohydrates are chemically defined as "polyhydroxy aldehyde or polyhydroxy
ketones or complex substances which on hydrolysis yield polyhydroxy aldehyde or
polyhydroxy ketone.”
Most of carbohydrates have formula:
(CH2O)n
Classification
1- Monosaccharides
Simplest carbohydrates, containing one sacchar unit. e.g. Glucose, Fructose, Galactose
2- Disaccharides
Disaccharides contain two sacchar units. e.g.:Sucrose (glucose + fructose)Lactose (glucose +
galactose)Maltose (glucose + glucose)
3- Oligosaccharides
These contain 3-10 monosaccharide units e.g. maltotriose
4- Polysaccharides
These contain more than 10 monosaccharide units. e.g.:Homopolysaccharides (Starch,
Glycogen, Cellulose, Chitin, Inulin)
Heteropolysaccharides (Glycosaminoglycans)
Chemical Properties of
Carbohydrates
Molisch Test: specific for carbohydrates.
1.
2. Iodine Test: presence of polysaccharides.

3. Benedict's Test: presence of reducing sugars.
4. Barfoed's Test: test used for detecting the presence of monosaccharides.
5. Seliwanoff's Test: distinguish between aldoses and ketoses.
Molisch test
This test is specific for all carbohydrates. Monosaccharide gives a rapid
positive test.
Objective:
To identify the carbohydrate from other macromolecules, lipids and proteins.
Principle
• The test reagent H2S04 dehydrates monosaccharides to form furfural or its
derivates.
• The furfural further react with α-naphthol present in the test reagent
(Molisch Reagent) to produce a purple ring.
Iodine test
polysaccharides.
Iodine test is a qualitative test for detection of
By this test we can also differentiate between starch, dextrin and glycogen.
Principle
This test depends upon the property of adsorption possessed by the large polysaccharide
molecules. Helically coiled polysaccharides adsorbs smaller molecules of iodine on their
surfaces to form a complex of ill defined chemical nature. At least eight monosaccharide
units should be joined in linear chain to give the colored complex.
Alkaline pH and increased temperature decrease the adsorption
property.
Effect Of pH: Perform iodine test and add 10% NaOH drop by
drop till the color disappears. Now add conc. Sulphuric acid
drop by drop till color reappears. INTERPRETATION: Iodine
test is positive only in acidic medium.
Effect Of Temperature: Perform iodine test and heat the test tube till
the color disappears. Now cool the test tube till color
reappears.
INTERPRETATION: Iodine test is negative at higher temperatures and positive
at lower temperatures. It is better to perform it at or near room temperature.
Benedict's Test
To detect the presence of reducing sugars.
• All monosaccharides are reducing sugars; they all have a free reactive
carbonyl group.
• Some disaccharides have exposed carbonyl groups and are also reducing
sugars. Other disaccharides such as sucrose are non-reducing sugars and will
not react with Benedict's solution
Principle
• The copper sulfate (CuSO4) present in Benedict's solution reacts with electrons from the
aldehyde or ketone group of the reducing sugar in alkaline medium.
• Reducing sugars are oxidized by the copper ion in solution to form a carboxylic acid and a
reddish precipitate of copper (I) oxide.
Barfoed’s Test
Objective : To distinguish if reducing sugar is a monosaccharide or disaccharides.

Principle
This test differs from Benedict’s test in aspect that the reduction of cupric ion is carried
out in a mildly acidic medium.
Since acidic medium is unfavorable for reduction, only strongly reducing carbohydrates
i.e. monosaccharides, reacts very fast and give positive result within 3 minutes.
Barfoed’s reagent consists of Copper acetate & Glacial acetic acid.

PRECAUTION: chloride ions interfere with the test therefore shouldn't be carried out with urine sample.
Selivanoff’s Test
Objective : To distinguish between ketose and aldose sugar.
Principle
Selivanoff’s reagent consists of
Resorcinol
Conc. HCL
When treated with HCL ketoses are dehydrated more quickly, giving a the furfural variant
(5-hydroxymethyl fufural) that is then condensed by the chemical resorcinol, resulting in
a cherry-red color compound. Aldoses react to create the identical product but with a
slower rate producing a faint pink to yellow color complex.
Precaution:
Final conc. Of HCL should not increase more than 12% in the reagent otherwise aldohexoses will convert to ketohexoses thus
it will give positive result.
Prolong boiling may also convert aldohexoses into ketohexoses. Hence boiling should be restricted to 30sec.

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Qualitative Analysis

2. Iodine Test: presence of polysaccharides.

Objective : To distinguish if reducing sugar is a monosaccharide or disaccharides.

Barfoed’s reagent consists of Copper acetate & Glacial acetic acid.

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